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通过与HRP酶催化和磁性纳米颗粒相结合的托脚辅助循环扩增法测定前列腺癌外泌体来源的miRNA。

Determination of miRNA derived from exosomes of prostate cancer via toehold-aided cyclic amplification combined with HRP enzyme catalysis and magnetic nanoparticles.

作者信息

Wang Qinjun, Liu Jingjian, Zeng Jiantao, Yang Zhiming, Ran Fengying, Wu Lun, Yang Guangyi, Mei Quanxi, Wang Xisheng, Chen Qinhua

机构信息

Department of Urology, Shenzhen Longhua District Central Hospital, Shenzhen, 518110, China.

Sinopharm Dongfeng General Hospital, Hubei University of Medicine, Shiyan, 442008, China.

出版信息

Anal Biochem. 2021 Oct 1;630:114336. doi: 10.1016/j.ab.2021.114336. Epub 2021 Aug 14.

Abstract

MicroRNAs (miRNAs) play a significant role in tumorigenesis and tumor development. Exosomal microRNA-141 (miRNA-141, miR-141) has been reported to be overexpressed in prostate cancer (PCa) and has become a potential biomarker for the diagnosis of PCa. Herein, a novel fluorescent biosensor based on toehold-aided cyclic amplification combined with horseradish peroxidase (HRP) enzyme catalysis and magnetic nanoparticles (MNPs) was designed for determination of the exosomes-derived microRNA-141 (miRNA-141, miR-141). The synergy of HRP enzyme catalysis and toehold mediated strand display reaction (TSDR) increase the sensitivity of the method, and the good separation ability of MNPs ensures the specificity of the method. Therefore, under the optimized experimental conditions, the highly sensitive and specific detection of miRNA-141 can be realized, and the detection limit is as low as 10 fM. More importantly, the biosensor successfully determinates the exosomal miR-141 in the plasma of patients with PCa.

摘要

微小RNA(miRNA)在肿瘤发生和肿瘤发展中发挥着重要作用。据报道,外泌体微小RNA-141(miRNA-141,miR-141)在前列腺癌(PCa)中过表达,并已成为PCa诊断的潜在生物标志物。在此,设计了一种基于托脚辅助循环扩增结合辣根过氧化物酶(HRP)酶催化和磁性纳米颗粒(MNP)的新型荧光生物传感器,用于测定外泌体来源的微小RNA-141(miRNA-141,miR-141)。HRP酶催化与托脚介导的链展示反应(TSDR)的协同作用提高了该方法的灵敏度,而MNP良好的分离能力确保了该方法的特异性。因此,在优化的实验条件下,可以实现对miRNA-141的高灵敏和特异性检测,检测限低至10 fM。更重要的是,该生物传感器成功测定了PCa患者血浆中的外泌体miR-141。

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